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Help: Subcloning

David Antonetti dantonetti at psghs.edu
Wed Jan 12 09:09:24 EST 2000


We are subcloning two separate PCR products for different cDNAs.  We
have ligated and transformed both inserts into 3 different vectors and
two
differenct e coli lines and get the same result.  We obtain amp
resistant colonies but we cannot retrieve any plasmid DNA using either
alkaline lysis
and phenol chloroform or using qiagen midi  kit.  However, if we try to
PCR from the plasmid prep we obtain product with the correct insert.
Other plasmids prepared in parrallel are fine.  Anyone with any
suggestions?

David Antonetti
dantonetti at psu.edu





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